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Antimicrobial and Antioxidant Efficacy of Acokanthera Oblongifolia Hochst

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Antimicrobial and Antioxidant Efficacy of Acokanthera Oblongifolia Hochst OPEN ACCESS International Journal of Pharmacology ISSN 1811-7775 DOI: 10.3923/ijp.2017.1086.1091 Research Article Antimicrobial and Antioxidant Efficacy of Acokanthera oblongifolia Hochst. (Apocynaceae) 1Wilfred Otang-Mbeng and 2Anthony Jide Afolayan 1School of Biology and Environmental Sciences, Faculty of Natural Sciences and Agriculture, University of Mpumalanga, Mbombela Campus, P/bag X11283, 1200 Nelspruit, Mpumalanga, South Africa 2Medicinal Plants and Economic Development (MPED) Research Centre, Department of Botany, University of Fort Hare, Private Bag X1314, 5700 Alice, South Africa Abstract Background and Objective: Acokanthera oblongifolia is an evergreen medicinal shrub used for snakebites, itches, wounds and internal worms and the relief of itchy conditions and other skin disorders by the Mpondo and Xhosa tribes in South Africa. The objective of this study was to investigate the efficacy of the plant extracts against selected pathogens that cause human skin disorders and evaluation of the antioxidant capability for validation of folk uses of the plant. Materials and Methods: The agar diffusion and micro-dilution methods were used to determine the antimicrobial activities of the extracts against selected bacteria and fungi. The data were subjected to one way analysis of variance (ANOVA) and Duncan’s Multiple Range Test (DMRT) was used to determine significant differences (p<0.05) among treatment means. Results: The highest antibacterial activity (inhibition zone diameter >19 mm) was obtained with the acetone extract against Pseudomonas aeruginosa, Shigella sonnei, Shigella flexneri, Bacillus cereus, Streptococcus pyogens and Bacillus subtilis; by the ethanol extract against B. cereus. None of the extracts was active against the tested fungi, apart from the acetone extract which showed strong inhibitory activity against Candida glabrata. The ethanol extract showed a higher ABTS scavenging than those of gallic acid and BHT at concentrations lower than 0.15 mg mLG1. Conclusion: The in vitro antibacterial activity of the acetone extract of A. oblongifolia against the tested pathogens has provide scientific evidence to justify the ethnomedicinal use of A. oblongofolia against skin disorders in the study area and also indicate that the plant is a potential source for the development of antimicrobial and antioxidant agents. Key words: Acokanthera oblongifolia, skin ailments, ethnomedicinal investigation, free-radical scavenging activity, antimicrobial activity Received: April 07, 2017 Accepted: June 06, 2017 Published: October 15, 2017 Citation: Wilfred Otang-Mbeng and Anthony Jide Afolayan, 2017. Antimicrobial and antioxidant efficacy of Acokanthera oblongifolia Hochst. (Apocynaceae). Int. J. Pharmacol., 13: 1086-1091. Corresponding Author: Wilfred Otang-Mbeng, School of Biology and Environmental Sciences, Faculty of Natural Sciences and Agriculture, University of Mpumalanga, Mbombela Campus, P/bag X11283, 1200 Nelspruit, Mpumalanga, South Africa Tel: +27 (0) 130020235 Copyright: © 2017 Wilfred Otang-Mbeng and Anthony Jide Afolayan. This is an open access article distributed under the terms of the creative commons attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited. Competing Interest: The authors have declared that no competing interest exists. Data Availability: All relevant data are within the paper and its supporting information files. Int. J. Pharmacol., 13 (8): 1086-1091, 2017 INTRODUCTION MATERIALS AND METHODS Medicinal plants have been used by mankind for the This study was carried out from December 2015 to management of diverse diseases since antiquity1. According February 2017 in the Amathole District, Eastern Cape, to the World Health Organisation, about 20,000 plant species South Africa. are used as medicines1,2. The widespread usage of medicinal plants in many cultures of the world could be attributed to a Extraction procedure: The leaves of A. oblogifolia were number of reasons including affordability and limited sterilized with 70% analytical grade ethanol and then washed availability of Western medicine as well as the popular believe with distilled water. Dried, pulverised leaf samples were that herbal medicines are less toxic since they are natural1. shaken for 24 h in ethanol and acetone within separate Natural products, whether standardised plant extracts or pure conical flasks placed on an orbital shaker. The solutions were compounds provide opportunities for the development of filtered through Whatman No. 1 filter paper in a Buchner new drug leads partly due to their unmatched chemical funnel and the filtrates were dried in a rotavapor at 40EC 1 diversity3,4. Natural products and related drugs are used to under reduced pressure. A 20 mg mLG stock solution was treat 87% of infections caused by bacteria, immunological formed by dissolving the extracts in their respective solvents disorders and cancer cases and about 25% of prescribed drugs of extraction. in the world originate from medicinal plants5,6. Microorganisms: Antibacterial studies were conducted Acokanthera oblongifolia Hochst. (Apocynaceae) is a using five Gram-positive bacteria (Staphylococcus aureus, common medicinal plant used for the treatment of various Streptococcus pyogenes, Enterococcus faecalis, Bacillus cereus diseases in the Eastern Cape Province of South Africa. It is an and Bacillus subtilis) and six Gram-negative bacteria ornamental shrub with densely clustered white flowers (Salmonella typhimurium, Escherichia coli, Pseudomonas located in leaf axils. The plant can grow to a height of 3 m as aeruginosa, Shigella sonnei, Shigella flexneri and Klebsiella a shrub and up to 6 m as a small tree. The dark-green leaves pneumonia). Antifungal activities were evaluated using are elliptic, opposite and leathery in texture. Because all parts Candida krusei and Candida glabrata. The choice of these of the plant are toxic, it has been used as arrow poison and microbes was attributed to their significance as pathogens related species in other parts of the world have been reported that cause human skin disorders. The microbes were American as causing deaths7. The ripe berries may cause salivation, Type Culture Collection (ATCC) purchased from Total vomiting and abdominal pain when ingested. The milky the Laboratory, South Africa. sap is irritant to the skin and eyes8. Hence the plant is also known as poison bush, bushman’s poison, or poison tree Antimicrobial susceptibility assays: Nutrient agar and because of its toxicity7. The leaf or wood decoction of the plant Sabouraud Dextrose Agar (SDA) media were prepared as per is used in local medicinal treatments for snakebites, itches, the recommendations of the manufacturer and poured into wounds and internal worms and an ointment made from sterilized disposable petri dishes under aseptic conditions. The the root scrapings of Acokanthera oblongifolia is used for labeled plates were inoculated with 100 µL of 0.5 Mcfarland the relief of itchy conditions and other skin disorders by the solutions of the respective organisms and loaded with extracts Mpondo and Xhosa tribes of the Eastern Cape Province of of A. oblongifolia (50 mg mLG1) into 6 mm wells. Positive 8 South Africa . In view of the importance of A. oblongifolia in controls for fungi and bacteria were nystatin and gentamycin ethnopharmacology and the dearth of scientific literature of respectively. The plates were incubated at 37ºC and the zones the in vitro medicinal potential of A. oblongifolia, the present of inhibition were recorded after 24 h. The Minimum Inhibitory study was undertaken to investigate the efficacy of the Concentration (MIC) of the plant extracts was determined acetone and ethanol extracts of the plant against selected using 96 well microtitre plates9. The least concentration of pathogens that cause human skin disorders and evaluation of plant extract that killed the microbes was considered as MIC. the antioxidant capability for validation of folk uses of the plant. This study has provided new evidence necessary for Assay of DPPH scavenging activity: The DPPH the scientific validation of the claimed medicinal potential of radical-scavenging activity of the test extracts was examined A. obolongifolia extracts and a basic understanding of the as previously described by Erukainure et al.10. Different extract plant‘s efficacy which may lend further support to the concentrations (0.025-0.5 :g mLG1) were added to equal widespread use of traditional medicine in health care systems volumes of methanolic solution of DPPH (100 :M) and in South Africa. the mixture was left for 30 min under darkness at room 1087 Int. J. Pharmacol., 13 (8): 1086-1091, 2017 temperature. Vitamin C and Rutin were used as standard Absorbance of the control- Absorbance of the sample (2) controls. Three replicates were made for each test sample. The ABTS+ scavenging activity (%) = 100 Absorbance (A) was measured after 30 min at 518 nm and the Absorbance of the control percentage antioxidant activity was calculated according to the Eq. 1: Reducing power: Different extracts concentrations (0.025-0.05 :g mLG1) prepared in distilled water were mixed Absorbance of the control- with 2.5 mL of 0.2 M phosphate buffer (pH 6.6) and 2.5 mL Absorbance of extract or standard (1) 4 DPPH scavenging activity (%) = 100 potassium ferricyanide (1% w/v) . The resulting mixture was Absorbance of the control incubated at 50EC for 20 min, followed by the addition of 2.5 mL
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